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Title: Cloning and expression of Bos indicus interleukin-4 in mammalian cells
Authors: Prashanth, T
Reddy, G R
Suryanaryana, V V S
Dechamma, H J
Keywords: Chinese hamster ovary cells;Cytokines;Dendritic cells;Interleukin-4;Nitric oxide
Issue Date: May-2013
Publisher: NISCAIR-CSIR, India
Abstract: Dendritic cells (DC) which are located at the interface of innate and adaptive immunity are targets of infection by many RNA and DNA viruses. Advances in the ex vivo generation of monocyte derived non proliferating dendritic cells have been used for clinical application like immunotherapy. IL-4 cytokine plays essential role in the maturation and generation of DCs. Bos indicus interleukin 4 (boIL-4) 408 bp was amplified from PBMC’s and cloned in pBSIIKS+ vector. The sequence analysis showed N terminal 69 bp signal sequence and one N-glycosylation site. The phylogenetic tree analysis showed that Bos indicus IL-4 is closely related to the ruminant IL-4 and least sharing of genetic line of human and mouse IL-4. The recombinant boIL-4 protein was expressed in CHO cells which secreted a 16 kDa protein which was confirmed by SDS PAGE and western blotting. The rec-boIL-4 protein proliferated the bovine PBMC’s, decreased production of nitric oxide in antigen stimulated macrophages, and phagocytosed the  micro particles confirming its activity on dendritic cells.
Page(s): 352-356
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.51(05) [May 2013]

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